A B S T R A C T The detection of platelet isoantibodies by the release of ['H]serotonin from platelets has been evaluated. The conditions for optimal release of ['H]serotonin with platelet isoantibodies using a microtechnique have been defined. A group of cardiac surgery patients were followed pre-and post-transfusions, with 48% developing a positive serotonin release assay. Of these patients, 16% also had a platelet complement-fixing and/or lymphocytotoxic isoantibody. There was variation in the degree of correlation between ['H]serotonin release and lymphocytotoxicity using individual National Institutes of Health typing serum. The matching obtained between family members by both techniques showed a close correlation when each technique was evaluated separately using the same NIH typing serum. The detection of isoantibodies in patients with hematological malignancies correlated with the unresponsiveness to unmatched platelet transfusions in 15 out of 17 cases. The use of the patient's isoantibody to matched platelets of family members by ['H]serotonin release correlated well with the clinical response to transfusion with these platelets. The data suggest that (a) platelet isoantibodies can be